"The Bubbler," a test you don't want to fail
Brown University
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| The Bubbler, a breathalyzer device that reverse-transcribes RNA from airborne SARS-CoV-2 in breath, predicts lower respiratory tract involvement and is less invasive than alternative testing approaches, researchers say. |
A team of researchers at Brown University and Rhode Island Hospital has developed a unique method for detecting SARS-CoV-2 in breath samples using viral RNA detection.
The researchers created a breathalyzer device that
reverse-transcribes RNA from airborne virus particles into DNA, which can then
be tested for COVID-19 via a common PCR test. The device, dubbed the Bubbler,
uses an enzymatic reaction mixture to detect viral RNA and convert it to DNA,
where it can be measured using a PCR instrument. The Bubbler can also barcode
that DNA, allowing a sample to be linked directly to the patient it came from
and be used for sequencing.
In a new study in the Journal of Molecular Diagnostics, the investigators report on the design and testing of the device.
“The Bubbler is more likely to be a better indicator of
current infection than nasopharyngeal swabs,” said lead investigator William G.
Fairbrother, a professor in the Department of Molecular Biology, Cell Biology
and Biochemistry at Brown.
The Bubbler is a glass tube with a glass pipette into which
patients exhale. The tube is filled with a reverse transcription reaction
mixture and cold mineral oil. The name of the device comes from the bubbling
sound that occurs when the patient exhales into the device.
In the study, 70 patients treated in the Emergency
Department at Rhode Island Hospital between May 2020 and January 2021 were
screened. Samples from three points in the respiratory tract were tested:
Saliva obtained from tongue scrapes in the mouth and samples of 15 seconds of
exhaled breath collected in the Bubbler were compared to a conventional
nasopharyngeal swab PCR test.
The study determined that SARS-CoV-2 can be readily detected
in the breath and is more predictive of lower respiratory tract involvement.
Viral RNA is more enriched in the breath relative to oral samples, while oral
samples include cells involved with SARS-CoV-2 replication that breath samples
do not. This suggests the viral signal detected in the Bubbler comes from
active viral particles, the authors said.
Bubbler kits were processed in a negative pressure bench top
box to prevent contamination of amplicon particles in the laboratory. (Photo:
William G. Fairbrother)
Tests for COVID-19 usually use samples collected from the
upper respiratory tract by saliva or nasopharyngeal swab. Positive samples
contain active virus, but viral load in the upper respiratory tract is not
correlated with symptoms in the lower respiratory tract, such as pneumonia.
“Involvement of the lower respiratory tract is often a
precursor to severe COVID-19, so there is an argument for a more direct
sampling focused on exhaled breath,” Fairbrother said.
Virus detection by the Bubbler is similar to a hospital-swab
PCR test. However, because swab tests detect viral RNA fragments in cells that
persist in previously infected cells, PCR tests can return a positive result
for months after infection. By contrast, the Bubbler detects airborne viral
particles and is a better measure of risk of contagion, the authors said. It
can also be adapted for environmental sampling in hospitals, transportation
hubs and closed environments like offices, ships and planes, the investigators
wrote in the study.
“Another advantage of the Bubbler is the barcoding, which
enables high-throughput RNA virus testing at a fraction of the cost of
conventional testing,” Fairbrother said.
The barcode returns a viral sequence that also supports strain identification, he said, which may prove useful as more information is learned about transmissibility and possible strain-specific treatment decisions.
It can also be used for simultaneous batches of pooled samples,
provides additional information such as viral load and strain identity, and
eliminates the need for stabilizing a sample, potentially allowing the assay to
be performed at home.
The investigators also demonstrated how the Bubbler might be adapted to detect virus in airborne samples. To model the movement of droplets exhaled in human breath, three unique nucleic acid samples were added to three personal humidifiers at different locations at varied distances from the Bubbler in a room with high airflow and a room with low airflow.
Although a
detailed exploration of this application was beyond the scope of the study, the
results demonstrate the potential to use aerosolized nucleic acids to
quantitatively map airflow in indoor spaces, and to detect SARS-CoV-2 in the
air.
“Such technology could be useful in restoring service to
industries such as hotels, cruise ships and casinos,” Fairbrother said. “There
is also an epidemiological benefit to routine testing of air at early warning
sites such as transportation hubs and hospital emergency departments.”
Other Brown University contributors to this study include
Chaorui Duan, Luke Buerer, Jing Wang, Gavin Sabalewski, Gregory D. Jay, Sean F.
Monaghan and Andrea E. Arena. The research was supported by National Institute
of General Medical Sciences grants R01GM105681 and R01GM127472, and the Roddy
Foundation grant GR5290840.